MF7000871 Rv3272, mutant D175A (Mycobacterium tuberculosis) 5yiy X-ray 2.50 Homodimer Mycobacterium tuberculosis 30342240 Karade SS, Pandey S, Ansari A, Das S, Tripathi S, Arora A, Chopra S, Pratap JV, Dasgupta A Biochim Biophys Acta Proteins Proteom 2019 3 1867 317-330 The availability of complete genome sequence of Mycobacterium tuberculosis has provided an important tool to understand the mycobacterial biology with respect to host-pathogen interaction, which is an unmet need of the hour owing to continuous increasing drug resistance. Hypothetical proteins are often an overlooked pool though half the genome encodes for such proteins of unknown function that could potentially play vital roles in mycobacterial biology. In this context, we report the structural and functional characterization of the hypothetical protein Rv3272. Sequence analysis classifies Rv3272 as a Family III CoA transferase with the classical two domain structure and conserved Aspartate residue (D175). The crystal structure of the wild type protein (2.2 Å) demonstrated the associated inter-locked dimer while that of the D175A mutant co-crystallized with octanoyl-CoA demonstrated relative movement between the two domains. Isothermal titration calorimetry studies indicate that Rv3272 binds to fatty acyl-CoAs of varying carbon chain lengths, with palmitoyl-CoA (C16:0) exhibiting maximum affinity. To determine the functional relevance of Rv3272 in mycobacterial biology, we ectopically expressed Rv3272 in M. smegmatis and assessed that its expression encodes significant alteration in cell surface with marked differences in triacylglycerol accumulation. Additionally, Rv3272 expression protects mycobacteria from acidic, oxidative and antibiotic stress under in vitro conditions. Taken together, these studies indicate a significant role for Rv3272 in host-pathogen interaction. GO:0016740 transferase activity 2 1 Homooligomeric enzymes Homodimeric enzymes All chains according to the most probable oligomerization state stored in PDBe were considered. A Probable fatty acyl-CoA transferase Rv3272 Mycobacterium tuberculosis P96877 5 394 98% MPTSNPAKPLDGFRVLDFTQNVAGPLAGQVLVDLGAEVIKVEAPGGEAARQITSVLPGRPPLATYFLPNNRGKKSVTVDLTTEQAKQQMLRLADTADVVLEAFRPGTMEKLGLGPDDLRSRNPNLIYARLTAYGGNGPHGSRPGIDLVVAAEAGMTTGMPTPEGKPQIIPFQLVDNASGHVLAQAVLAALLHRERNGVADVVQVAMYDVAVGLQANQLMMHLNRAASDQPKPEPAPKAKRRKGVGFATQPSDAFRTADGYIVISAYVPKHWQKLCYLIGRPDLVEDQRFAEQRSRSINYAELTAELELALASKTATEWVQLLQANGLMACLAHTWKQVVDTPLFAENDLTLEVGRGADTITVIRTPARYASFRAVVTDPPPTAGEHNAVFLARP 394 secondary structure helix 22 34 secondary structure helix 47 51 secondary structure helix 66 70 secondary structure helix 82 94 secondary structure helix 106 111 secondary structure helix 114 122 secondary structure helix 145 153 secondary structure helix 172 197 secondary structure helix 206 214 secondary structure helix 214 223 secondary structure helix 267 278 secondary structure helix 280 284 secondary structure helix 291 298 secondary structure helix 298 311 secondary structure helix 314 325 secondary structure helix 334 339 secondary structure helix 341 347 secondary structure helix 386 391 secondary structure strand 14 17 secondary structure strand 37 42 secondary structure strand 74 77 secondary structure strand 98 101 secondary structure strand 126 131 secondary structure strand 201 205 secondary structure strand 251 255 secondary structure strand 260 264 secondary structure strand 328 330 secondary structure strand 350 355 secondary structure strand 358 363 secondary structure strand 368 369 pfam PF02515 CoA-transferase family III 10 387 B Probable fatty acyl-CoA transferase Rv3272 Mycobacterium tuberculosis P96877 5 393 98% MPTSNPAKPLDGFRVLDFTQNVAGPLAGQVLVDLGAEVIKVEAPGGEAARQITSVLPGRPPLATYFLPNNRGKKSVTVDLTTEQAKQQMLRLADTADVVLEAFRPGTMEKLGLGPDDLRSRNPNLIYARLTAYGGNGPHGSRPGIDLVVAAEAGMTTGMPTPEGKPQIIPFQLVDNASGHVLAQAVLAALLHRERNGVADVVQVAMYDVAVGLQANQLMMHLNRAASDQPKPEPAPKAKRRKGVGFATQPSDAFRTADGYIVISAYVPKHWQKLCYLIGRPDLVEDQRFAEQRSRSINYAELTAELELALASKTATEWVQLLQANGLMACLAHTWKQVVDTPLFAENDLTLEVGRGADTITVIRTPARYASFRAVVTDPPPTAGEHNAVFLARP 394 secondary structure helix 22 34 secondary structure helix 47 51 secondary structure helix 66 70 secondary structure helix 82 94 secondary structure helix 106 111 secondary structure helix 114 122 secondary structure helix 145 153 secondary structure helix 172 197 secondary structure helix 206 214 secondary structure helix 214 222 secondary structure helix 267 278 secondary structure helix 280 284 secondary structure helix 291 298 secondary structure helix 298 310 secondary structure helix 311 313 secondary structure helix 314 325 secondary structure helix 334 339 secondary structure helix 341 347 secondary structure helix 386 391 secondary structure strand 14 17 secondary structure strand 37 42 secondary structure strand 74 77 secondary structure strand 98 101 secondary structure strand 126 131 secondary structure strand 201 205 secondary structure strand 251 255 secondary structure strand 260 264 secondary structure strand 328 331 secondary structure strand 350 353 secondary structure strand 360 363 secondary structure strand 368 369 pfam PF02515 CoA-transferase family III 10 387 Indirect evidence The full structure participates in mutual synergistic folding. CoA-transferase family III The structure of CaiB reveals a spectacular fold where two monomers are interlaced to form an interlocked dimer with large interface (PMID:15518548). The folding path of CaiB and other proteins of this fold must be quite complex because the dimer cannot be formed by the individually folded monomers, partial unfolding would have to take place to break the CaiB dimer, thus it is most probably very stable (PMID:15518548). A N/A B N/A MF7000131 MF7000132 MF7000133 MF7000134 MF7000135 MF7000871